Department of Physiological Sciences, Eastern Virginia Medical School, Norfolk, VA, USA.
Department of Obstetrics and Gynecology, University of Kentucky, Lexington, KY, USA.
FASEB J. 2021 Apr;35(4):e21481. doi: 10.1096/fj.202002547RR.
The midcycle luteinizing hormone (LH) surge initiates a cascade of events within the ovarian follicle which culminates in ovulation. Only mural granulosa cells and theca cells express large numbers of LH receptors, and LH-stimulated paracrine mediators communicate the ovulatory signal within the follicle. Recent reports identified the neuropeptide neurotensin (NTS) as a product of granulosa cells. Here, we demonstrate that granulosa cells were the primary site of NTS expression in macaque ovulatory follicles. Granulosa cell NTS mRNA and protein increased after human chorionic gonadotropin (hCG) administration, which substitutes for the LH surge. To identify ovulatory actions of NTS, a NTS-neutralizing antibody was injected into preovulatory macaque follicles. hCG administration immediately followed, and ovaries were removed 48 hours later to evaluate ovulatory events. Follicles injected with control IgG ovulated normally. In contrast, 75% of NTS antibody-injected follicles failed to ovulate, containing oocytes trapped within unruptured, hemorrhagic follicles. Serum progesterone was unchanged. Of the three NTS receptors, SORT1 was highly expressed in follicular granulosa, theca, and endothelial cells; NTSR1 and NTSR2 were expressed at lower levels. Excessive blood cells in NTS antibody-injected follicles indicated vascular anomalies, so the response of monkey ovarian endothelial cells to NTS was evaluated in vitro. NTS stimulated endothelial cell migration and capillary sprout formation, consistent with a role for NTS in vascular remodeling associated with ovulation. In summary, we identified NTS as a possible paracrine mediator of ovulation. Further investigation of the NTS synthesis/response pathway may lead to improved treatments for infertility and novel targets for contraception.
黄体中期的促黄体激素 (LH) 激增引发了卵巢卵泡内的一系列事件,最终导致排卵。只有壁层颗粒细胞和膜细胞表达大量的 LH 受体,LH 刺激的旁分泌介质在卵泡内传递排卵信号。最近的报告将神经肽神经降压素 (NTS) 鉴定为颗粒细胞的产物。在这里,我们证明了 NTS 是猕猴排卵卵泡中颗粒细胞的主要表达部位。人绒毛膜促性腺激素 (hCG) 给药后,颗粒细胞 NTS mRNA 和蛋白增加,这替代了 LH 激增。为了确定 NTS 的排卵作用,将 NTS 中和抗体注入预排卵的猕猴卵泡中。hCG 立即给药,48 小时后取出卵巢以评估排卵事件。用对照 IgG 注射的卵泡正常排卵。相比之下,75%的 NTS 抗体注射的卵泡未能排卵,含有被困在未破裂、出血卵泡内的卵母细胞。血清孕激素没有变化。在三种 NTS 受体中,SORT1 在卵泡颗粒细胞、膜细胞和内皮细胞中高度表达;NTSR1 和 NTSR2 表达水平较低。NTS 抗体注射的卵泡中过多的血细胞表明存在血管异常,因此评估了猴子卵巢内皮细胞对 NTS 的反应。NTS 刺激内皮细胞迁移和毛细血管芽形成,表明 NTS 在与排卵相关的血管重塑中起作用。总之,我们确定 NTS 是排卵的一种可能的旁分泌介质。对 NTS 合成/反应途径的进一步研究可能会导致改善不孕不育的治疗方法和新的避孕目标。